The antigen for Hep Par 1 antibody is the urea cycle enzyme carbamoyl phosphate synthetase 1
Hepatocyte paraffin 1 (Hep Par 1), a murine monoclonal antibody, is widely used in surgical pathology practice to determine the hepatocellular origin of neoplasms. However, identity of the antigen for Hep Par 1 is unknown. The aim of this study was to characterize the Hep Par 1 antigen. To identify...
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| Veröffentlicht in: | Laboratory investigation 2008-01, Vol.88 (1), p.78-88 |
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| creator | Butler, Samantha L Dong, Huijia Cardona, Diana Jia, Minghong Zheng, Ran Zhu, Haizhen Crawford, James M Liu, Chen |
| description | Hepatocyte paraffin 1 (Hep Par 1), a murine monoclonal antibody, is widely used in surgical pathology practice to determine the hepatocellular origin of neoplasms. However, identity of the antigen for Hep Par 1 is unknown. The aim of this study was to characterize the Hep Par 1 antigen. To identify the antigen, immunoprecipitation was used to isolate the protein from human liver tissue, and a distinct protein band was detected at approximately 165 kDa. The protein band was also present in small intestinal tissue, but was not present in several other non-liver tissues nor in three human hepatocellular carcinoma cell lines, Huh-7, HepG2, and LH86. The protein was purified and analyzed by mass spectrometry. It was identified as carbamoyl phosphate synthetase 1 (CPS1). CPS1 is a rate-limiting enzyme in urea cycle and is located in mitochondria. We demonstrated that hepatoid tumors (gastric and yolk sac) were immunoreactive with both Hep Par 1 antibody and anti-CPS1 antibody, further confirming the results of mass spectrometric analysis. We found that the three human hepatocellular carcinoma cell lines do not express either CPS1 RNA or protein. We confirmed that the gene was present in these cell lines, suggesting that suppression of CPS1 expression occurs at the transcriptional level. This finding may have relevance to liver carcinogenesis, since poorly differentiated hepatocellular carcinomas exhibit poor to absent immunoreactivity to Hep Par 1. In conclusion, we have identified the antigen for Hep Par 1 antibody as a urea cycle enzyme CPS1. Our results should encourage further investigation of potential role that CPS1 expression plays in liver pathobiology and carcinogenesis. |
| doi_str_mv | 10.1038/labinvest.3700699 |
| format | Article |
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However, identity of the antigen for Hep Par 1 is unknown. The aim of this study was to characterize the Hep Par 1 antigen. To identify the antigen, immunoprecipitation was used to isolate the protein from human liver tissue, and a distinct protein band was detected at approximately 165 kDa. The protein band was also present in small intestinal tissue, but was not present in several other non-liver tissues nor in three human hepatocellular carcinoma cell lines, Huh-7, HepG2, and LH86. The protein was purified and analyzed by mass spectrometry. It was identified as carbamoyl phosphate synthetase 1 (CPS1). CPS1 is a rate-limiting enzyme in urea cycle and is located in mitochondria. We demonstrated that hepatoid tumors (gastric and yolk sac) were immunoreactive with both Hep Par 1 antibody and anti-CPS1 antibody, further confirming the results of mass spectrometric analysis. We found that the three human hepatocellular carcinoma cell lines do not express either CPS1 RNA or protein. We confirmed that the gene was present in these cell lines, suggesting that suppression of CPS1 expression occurs at the transcriptional level. This finding may have relevance to liver carcinogenesis, since poorly differentiated hepatocellular carcinomas exhibit poor to absent immunoreactivity to Hep Par 1. In conclusion, we have identified the antigen for Hep Par 1 antibody as a urea cycle enzyme CPS1. Our results should encourage further investigation of potential role that CPS1 expression plays in liver pathobiology and carcinogenesis.</description><identifier>ISSN: 0023-6837</identifier><identifier>EISSN: 1530-0307</identifier><identifier>DOI: 10.1038/labinvest.3700699</identifier><identifier>PMID: 18026163</identifier><identifier>CODEN: LAINAW</identifier><language>eng</language><publisher>New York: Elsevier Inc</publisher><subject>Antibodies, Monoclonal - immunology ; Antigens - immunology ; Base Sequence ; Biological and medical sciences ; Biotechnology ; Blotting, Western ; Carbamoyl-Phosphate Synthase (Ammonia) - immunology ; Chromatography, Liquid ; CPS1 ; DNA Primers ; Female ; Fundamental and applied biological sciences. Psychology ; Hep Par 1 antibody ; Humans ; Immunoprecipitation ; Investigative techniques, diagnostic techniques (general aspects) ; Laboratory Medicine ; liver ; liver cancer ; Liver Neoplasms - enzymology ; Male ; Medical sciences ; Medicine ; Medicine & Public Health ; Pathology ; research-article ; Reverse Transcriptase Polymerase Chain Reaction ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry ; urea cycle</subject><ispartof>Laboratory investigation, 2008-01, Vol.88 (1), p.78-88</ispartof><rights>2008 United States & Canadian Academy of Pathology</rights><rights>United States and Canadian Academy of Pathology, Inc. 2008</rights><rights>2008 INIST-CNRS</rights><rights>Copyright Nature Publishing Group Jan 2008</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c582t-96c3aeb6aebed737528a8c4d65ec96e8c2ffe1b6804f7c704ae018ac8257830b3</citedby><cites>FETCH-LOGICAL-c582t-96c3aeb6aebed737528a8c4d65ec96e8c2ffe1b6804f7c704ae018ac8257830b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,4024,27923,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=20010164$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/18026163$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Butler, Samantha L</creatorcontrib><creatorcontrib>Dong, Huijia</creatorcontrib><creatorcontrib>Cardona, Diana</creatorcontrib><creatorcontrib>Jia, Minghong</creatorcontrib><creatorcontrib>Zheng, Ran</creatorcontrib><creatorcontrib>Zhu, Haizhen</creatorcontrib><creatorcontrib>Crawford, James M</creatorcontrib><creatorcontrib>Liu, Chen</creatorcontrib><title>The antigen for Hep Par 1 antibody is the urea cycle enzyme carbamoyl phosphate synthetase 1</title><title>Laboratory investigation</title><addtitle>Lab Invest</addtitle><addtitle>Lab Invest</addtitle><description>Hepatocyte paraffin 1 (Hep Par 1), a murine monoclonal antibody, is widely used in surgical pathology practice to determine the hepatocellular origin of neoplasms. However, identity of the antigen for Hep Par 1 is unknown. The aim of this study was to characterize the Hep Par 1 antigen. To identify the antigen, immunoprecipitation was used to isolate the protein from human liver tissue, and a distinct protein band was detected at approximately 165 kDa. The protein band was also present in small intestinal tissue, but was not present in several other non-liver tissues nor in three human hepatocellular carcinoma cell lines, Huh-7, HepG2, and LH86. The protein was purified and analyzed by mass spectrometry. It was identified as carbamoyl phosphate synthetase 1 (CPS1). CPS1 is a rate-limiting enzyme in urea cycle and is located in mitochondria. We demonstrated that hepatoid tumors (gastric and yolk sac) were immunoreactive with both Hep Par 1 antibody and anti-CPS1 antibody, further confirming the results of mass spectrometric analysis. We found that the three human hepatocellular carcinoma cell lines do not express either CPS1 RNA or protein. We confirmed that the gene was present in these cell lines, suggesting that suppression of CPS1 expression occurs at the transcriptional level. This finding may have relevance to liver carcinogenesis, since poorly differentiated hepatocellular carcinomas exhibit poor to absent immunoreactivity to Hep Par 1. In conclusion, we have identified the antigen for Hep Par 1 antibody as a urea cycle enzyme CPS1. Our results should encourage further investigation of potential role that CPS1 expression plays in liver pathobiology and carcinogenesis.</description><subject>Antibodies, Monoclonal - immunology</subject><subject>Antigens - immunology</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Blotting, Western</subject><subject>Carbamoyl-Phosphate Synthase (Ammonia) - immunology</subject><subject>Chromatography, Liquid</subject><subject>CPS1</subject><subject>DNA Primers</subject><subject>Female</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Hep Par 1 antibody</subject><subject>Humans</subject><subject>Immunoprecipitation</subject><subject>Investigative techniques, diagnostic techniques (general aspects)</subject><subject>Laboratory Medicine</subject><subject>liver</subject><subject>liver cancer</subject><subject>Liver Neoplasms - enzymology</subject><subject>Male</subject><subject>Medical sciences</subject><subject>Medicine</subject><subject>Medicine & Public Health</subject><subject>Pathology</subject><subject>research-article</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>Spectrometry, Mass, Electrospray Ionization</subject><subject>Tandem Mass Spectrometry</subject><subject>urea cycle</subject><issn>0023-6837</issn><issn>1530-0307</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp9kEGL1DAUx4Mo7uzqB_CgBGGPXV-SNsngSRZ1hQU9rDehvKavO1k6aU06C_XTG22Zue0hBPL_vf8LP8beCLgSoOyHHhsfHilNV8oA6O32GduISkEBCsxztgGQqtBWmTN2ntIDgChLXb1kZ8KC1EKrDft1tyOOYfL3FHg3RH5DI_-BkYv_r83QztwnPmXqEAm5m11PnMKfeU_cYWxwP8w9H3dDGnc4EU9zyPCEibh4xV502Cd6vd4X7OeXz3fXN8Xt96_frj_dFq6yciq22imkRudDrVGmkhatK1tdkdtqsk52HYlGWyg74wyUSCAsOisrYxU06oK9X3rHOPw-ZB_1w3CIIa-spQRpjCwhQ2KBXBxSitTVY_R7jHMtoP6nsz7qrFedeebdWnxo9tSeJlZ_GbhcAUwO-y5icD4dOZmVg9Bl5uTCpRyFe4qnHz61_e0yFHDK8o-tp_zjklN2--hzaXKegqPWR3JT3Q7-ifa_voevSQ</recordid><startdate>200801</startdate><enddate>200801</enddate><creator>Butler, Samantha L</creator><creator>Dong, Huijia</creator><creator>Cardona, Diana</creator><creator>Jia, Minghong</creator><creator>Zheng, Ran</creator><creator>Zhu, Haizhen</creator><creator>Crawford, James M</creator><creator>Liu, Chen</creator><general>Elsevier Inc</general><general>Nature Publishing Group US</general><general>Nature Publishing</general><general>Nature Publishing Group</general><scope>6I.</scope><scope>AAFTH</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7T5</scope><scope>7T7</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope></search><sort><creationdate>200801</creationdate><title>The antigen for Hep Par 1 antibody is the urea cycle enzyme carbamoyl phosphate synthetase 1</title><author>Butler, Samantha L ; Dong, Huijia ; Cardona, Diana ; Jia, Minghong ; Zheng, Ran ; Zhu, Haizhen ; Crawford, James M ; Liu, Chen</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c582t-96c3aeb6aebed737528a8c4d65ec96e8c2ffe1b6804f7c704ae018ac8257830b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><topic>Antibodies, Monoclonal - immunology</topic><topic>Antigens - immunology</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Blotting, Western</topic><topic>Carbamoyl-Phosphate Synthase (Ammonia) - immunology</topic><topic>Chromatography, Liquid</topic><topic>CPS1</topic><topic>DNA Primers</topic><topic>Female</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Hep Par 1 antibody</topic><topic>Humans</topic><topic>Immunoprecipitation</topic><topic>Investigative techniques, diagnostic techniques (general aspects)</topic><topic>Laboratory Medicine</topic><topic>liver</topic><topic>liver cancer</topic><topic>Liver Neoplasms - enzymology</topic><topic>Male</topic><topic>Medical sciences</topic><topic>Medicine</topic><topic>Medicine & Public Health</topic><topic>Pathology</topic><topic>research-article</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>Spectrometry, Mass, Electrospray Ionization</topic><topic>Tandem Mass Spectrometry</topic><topic>urea cycle</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Butler, Samantha L</creatorcontrib><creatorcontrib>Dong, Huijia</creatorcontrib><creatorcontrib>Cardona, Diana</creatorcontrib><creatorcontrib>Jia, Minghong</creatorcontrib><creatorcontrib>Zheng, Ran</creatorcontrib><creatorcontrib>Zhu, 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paraffin 1 (Hep Par 1), a murine monoclonal antibody, is widely used in surgical pathology practice to determine the hepatocellular origin of neoplasms. However, identity of the antigen for Hep Par 1 is unknown. The aim of this study was to characterize the Hep Par 1 antigen. To identify the antigen, immunoprecipitation was used to isolate the protein from human liver tissue, and a distinct protein band was detected at approximately 165 kDa. The protein band was also present in small intestinal tissue, but was not present in several other non-liver tissues nor in three human hepatocellular carcinoma cell lines, Huh-7, HepG2, and LH86. The protein was purified and analyzed by mass spectrometry. It was identified as carbamoyl phosphate synthetase 1 (CPS1). CPS1 is a rate-limiting enzyme in urea cycle and is located in mitochondria. We demonstrated that hepatoid tumors (gastric and yolk sac) were immunoreactive with both Hep Par 1 antibody and anti-CPS1 antibody, further confirming the results of mass spectrometric analysis. We found that the three human hepatocellular carcinoma cell lines do not express either CPS1 RNA or protein. We confirmed that the gene was present in these cell lines, suggesting that suppression of CPS1 expression occurs at the transcriptional level. This finding may have relevance to liver carcinogenesis, since poorly differentiated hepatocellular carcinomas exhibit poor to absent immunoreactivity to Hep Par 1. In conclusion, we have identified the antigen for Hep Par 1 antibody as a urea cycle enzyme CPS1. Our results should encourage further investigation of potential role that CPS1 expression plays in liver pathobiology and carcinogenesis.</abstract><cop>New York</cop><pub>Elsevier Inc</pub><pmid>18026163</pmid><doi>10.1038/labinvest.3700699</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Antibodies, Monoclonal - immunology Antigens - immunology Base Sequence Biological and medical sciences Biotechnology Blotting, Western Carbamoyl-Phosphate Synthase (Ammonia) - immunology Chromatography, Liquid CPS1 DNA Primers Female Fundamental and applied biological sciences. Psychology Hep Par 1 antibody Humans Immunoprecipitation Investigative techniques, diagnostic techniques (general aspects) Laboratory Medicine liver liver cancer Liver Neoplasms - enzymology Male Medical sciences Medicine Medicine & Public Health Pathology research-article Reverse Transcriptase Polymerase Chain Reaction Spectrometry, Mass, Electrospray Ionization Tandem Mass Spectrometry urea cycle |
| title | The antigen for Hep Par 1 antibody is the urea cycle enzyme carbamoyl phosphate synthetase 1 |
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